1a.Objectives (from AD-416):
The proposed work will provide basic and applied information for the development and transfer of appropriate and environmentally acceptable technologies for control of fruit flies and other invasive species. The major objectives of our projects are:.1)Study population dynamics and pest interactions with their natural enemies, host plants, and other pests in the ecosystem; 2)Identify attractants from host and non-host plants and determine physiological and environmental factors affecting or modulating pest behavior; 3)Improve attractants and trapping systems for surveillance, detection, and control of fruit flies and/or other tropical plant pests of quarantine significance; 4)Assess the efficacy and quality of laboratory-reared insects used in SIT and natural enemies for control of fruit flies and other tropical plant pests of quarantine significance, and determine factors limiting their effectiveness;.5)Develop basic understanding of the oriental fruit fly genome, annotation of functional proteins that regulate development, metabolism, sensory reception and sex determination; and 6)Develop area-wide IPM systems including integration of environmentally friendly replacements for organophosphate chemicals to reduce the economic impact of fruit flies and other tropical plant pests.

1b.Approach (from AD-416):
Test more environmentally acceptable pesticides and compounds for use in control or eradication program for fruit flies. Investigate genomics, biology, and behavior of fruit flies and their parasitoids. Evaluate augmentative parasite releases, mass trapping, male annihilation, sterile fly releases, novel insecticides and food bait sprays as control, suppression, or eradication technologies. Develop new or improved fruit fly strains, lures, attractants, parapheromones, and baits to be used in improved trap detection devices and/or current control systems. Investigate fruit fly and parasitoid olfactory, gustatory, and oviposition aensilla and their electrophysiology. Use genomics, computational biology and area-wide control for fruit flies and other pests.

3.Progress Report:
Continuing progress is being made on basic studies and development of area-wide control strategies for invasive pests of economic importance such as fruit flies. During the past year an annotated world bibliography of the host fruits of Malaysian fruit fly was completed and a searchable database of the host plants of the Mediterranean fruit fly was made available online. Volatile semiochemicals have been identified from insect host plants from fruit flies, coffee berry borer,and coconut rhinoceros beetle. These compounds are being tested for behavioral activity in both sensory and whole animal assays. A compound that may be attractive to Queensland fruit fly is being evaluated in Australia. An improved sprayable biopesticide male annihilation technique formulation was tested at the orchard level in Hawaii, Pakistan, and Brazil. Rapid reductions in male oriental, peach, and carambola fruit flies, respectively, occurred in all three demonstration trials. Results suggest SPLAT® MAT Spinosad ME (aka STATICTMME-spinosad) “attract and kill” formulations containing spinosad are more convenient and safer to handle than current liquid insecticide formulations which utilize more toxic and less desirable organophosphate insecticides. In addition, novel trilure solid lure and insecticide dispensers have been shown to be as effective as standard liquid lure formulations currently used on the U.S. mainland. Trials are now underway in California for use in detection and eradication of annual accidental introductions of fruit flies. Natural enemies of fruit flies have been successfully mass-reared and sent to both Brazil, Senegal and Tahiti through informal cooperative programs. The natural enemy, has been successfully established in Tahiti. Studies on the factors that limit the abundance of these natural enemies are being conducted at Pacific Basin Agricultural Research Center (PBARC) and in Brazil through laboratory studies. The use of sterile Mediterranean fruit flies as carriers for pheromone of the light brown apple moth was tested in a novel form of mating disruption termed mobile mating disruption. Small scale field tests demonstrated that flies fed moth pheromone could disrupt mating for a short time period. ARS scientists at PBARC also have identified differentially expressed proteins/genes specific to radiation treatment and nutritional deficiency at all ages of adult and pupa stages in oriental fruit fly. Manipulation of these target proteins/genes in fruit fly life stages and/or ages using RNA interference technology may help improve the quality of the fruit fly control program, especially with the Sterile Insect Technique (SIT). The genome of the oriental fruit fly has been sequenced and assembly and annotation are progressing rapidly.

4.Accomplishments
1.
Completion of an annotated world bibliography of the host fruits of Bactrocera latifrons. Bactrocera latifrons (Hendel) is a tephritid fruit fly of primarily Asian distribution, but its range has expanded through introductions into Hawaii, Japan, Tanzania, and Kenya, and poses a risk of introduction into other countries where it does not presently exist, including the U.S. Mainland. Fruits and vegetables in danger of attack are primarily in the tomato family (Solanaceae) and the squash family (Cucurbitaceae). Introduction of B. latifrons can have significant economic consequences, including damage and loss of food production, as well as requirements for inspection of imported susceptible commodities and for implementation of costly quarantine treatments to permit export of commodities susceptible to infestation by B. latifrons. An ARS scientist in Hilo, Hawaii, in collaboration with an APHIS pest risk analyst in Honolulu, Hawaii, prepared a summary of all host plants of B. latifrons reported in worldwide literature, with annotations on reported laboratory and field infestation data. This summary, now published and available online, will permit quarantine regulatory officials, both in the U.S. and in other countries, to better assess the risk of introduction of B. latifrons in imported or exported fruits and vegetables.

2.
Searchable database of the host plants of the Mediterranean fruit fly made available online. Through collaboration of USDA, ARS, Pacific Basin Agricultural Research Center (PBARC), Hilo, Hawaii, PUSDA-APHIS-PPQ-CPHST, Honolulu, Hawaii, and the Center for Integrated Pest Management (CIPM), Raleigh, North Carolina, a searchable summary of the host plants of the Mediterranean fruit fly (Medfly) was made available online in support of regulatory concerns for interstate and international commerce. The Medfly host plant database is a primary reference for regulatory officials for assessment of risk and for the development of mitigation protocols for Mediterranean fruit fly in interstate and international movements of fruit and vegetable commodities. The online publication for Medfly extends access to the host database. The online searchable database permits quarantine regulatory officials, both in the U.S. and in other countries, to better assess the risk of introduction of Mediterranean fruit fly in imported or exported fruits and vegetables.

3.
New male annihilation technique (MAT) biopesticide formulation shown to be effective against three different species of Bactrocera in three different countries. A new biopesticide (SPLAT®MAT Spinosad ME) was evaluated by Pacific Basin Agricultural Center (PBARC), Hilo, Hawaii, scientists and industry cooperators in demonstration trials as a male annihilation treatment against oriental fruit fly in Hawaii, peach fruit fly in Pakistan, and carambola fruit fly in Brazil. In Hawaii, field demonstration tests were conducted in commercial papaya orchards. Evaluations in Brazil and Pakistan compared standard methods (organophosphate insecticides) with one application of SPLAT® MAT Spinosad ME. Results also suggest a reduction of female flies over time. Our current studies suggest SPLAT® MAT Spinosad ME “attract and kill” formulations containing spinosad are more convenient and safer to handle than current liquid insecticide formulations which utilize more toxic and less desirable organophosphate insecticides.

4.
Proteomic approach to understanding fruit fly metabolic pathways. Insects such as the oriental fruit fly face many challenges during their lifetimes such as nutrition and other environmental factors. To cope with stresses, insects have developed various behavioral, biochemical, and physiological mechanisms. These responses can be detected at the proteomic level. ARS scientists at Pacific Basin Agricultural Center (PBARC) in Hilo, Hawaii, have identified differentially expressed proteins/genes specific to radiation treatment and nutritional deficiency at all ages of adult and pupa stages in oriental fruit fly. Manipulation of these target proteins/genes in fruit fly life stages and/or ages using RNA interference technology may help to improve on fruit fly quality for control program especially in Sterile Insect Technique (SIT).

5.
New trilure fruit fly trap dispensers being evaluated in California. Trilure fruit fly dispensers (TMR) that contain the three parapheromone lures trimedlure (TML), raspberry ketone (RK), and methyl eugenol (ME) are being evaluated in five citrus growing areas in California by UC and ARS,USDA Hawaii scientists. New outbreaks of invasive fruit flies (Diptera: Tephritidae) continue to threaten agriculture in California. Establishment of these pests often results in serious economic and environmental consequences associated with quarantine, control, and eradication programs. Currently California deploys 30,000 sets of individual traps with TML, ME, and RK (C-L) for fruit fly detection. TMR wafers have the potential to significantly reduce the cost of tephritid fruit fly detection trapping in California by providing effective detection capabilities for fruit flies such as Mediterranean fruit fly, oriental fruit fly and melon fly in a single trap, rather than needing to deploy separate traps (TML, ME and RK (C-L)) for each of the three species, representative of the three major fruit fly lure responding groups.

6.
Development of novel methods for studying fruit fly movement and responses to lures. An apparatus that detects Radio-Frequency Identification (RFID)-tagged fruit flies has been developed and tested in a field enclosure. This apparatus is specially designed to allow tagged flies to land only on areas where they can be detected. Work is ongoing to use RFID tagged fruit flies to analyse individual-level responses to lures under semi-natural conditions. In addition, automatic vision-based detection and quantification was used to create a high-resolution dataset representing the response of colony-reared melon flies to cuelure (insect pheromone) at different times of the day. Computer vision method and results with melon fly have been published (Manoukis and Jang 2013), and the same method has been employed in a study on oriental fruit fly responses to methly eugenol and basil oil.

7.
Genome sequencing of the oriental fruit fly. There is very little genetic information available on pest fruit fly species in the genus Bactrocera. Improving knowledge of Bactrocera at a genetic level can lead to the development of novel approaches towards monitoring and control of this genus. In the last year, research was focused at making initial releases of the oriental fruit fly genomic and transcriptomic databases. The oriental fruit fly RNA-Seq based transcriptome has been released as National Center for Biotechnology Information (NCBI) Bioproject PRJNA198716, including stage specific expression analysis, structural and functional annotation. The draft assembly of the genome was released as NCBI bioproject PRJNA208397, including complete structural and functional annotation for the initial release of the genome. In addition, analysis and annotation of the medfly genome was performed at USDA, ARS, Pacific Basin Agricultural Center (PBARC), Hilo, Hawaii, which was contributed to the medfly genome release associated with the i5K Insect and other Arthropod Genome Sequencing Initiative under NCBI bioproject PRJNA168120.